Abstract: The merits and disadvantages of thin-film rehydration-ultrasonic method and ethanol injection-DHPM were evaluated by comparing the physicochemical properties of fish oil(FO) nano liposomes prepared under individual optimum conditions.(liposome I and liposome II,respectively) The polydisperse index,particle size,zeta potential,transmission electron microscopy and p H of liposomes was detected. Result:Nanoliposomes Ⅱexhibited small eraverage particle,PDI and zeta potentialsize,than nano liposomes I. TEM and AFM analysis revealed that the particle integrity of liposome II was better than liposomes I. Conclusion :Compared with nanoliposomes I,nanoliposomes Ⅱ exhibites greater characteristies,and ethanol injection-DHPM could be a potent technique for the preparation of nanoliposomes.